Concern about the adequate supply of natural products for preclinical and clinical studies is a critical issue in drug development strategies. Efforts to obtain sufficient material for pre-clinical and clinical evaluation of natural products has demonstrated that bulk collections from natural populations are neither economically nor ecologically feasible. Advances being made in invertebrate cell culture and culture of marine microorganisms may provide viable alternative sources of bioactive compounds with pharmaceutical potential. The objectives of the proposed research are to determine the biogenetic origin of bioactive marine natural products at the cellular level within selected deep water sponges, to address the issues of host vs. associated microbe production of metabolites, and to supply such bioactive metabolites for drug development strategies. The Specific Aims of this proposal are: 1) To determine the biogenetic origin of bioactive metabolite(s) isolated from selected deep water marine sponges using dissociation, separation and detection techniques including density gradient fractionation, differential centrifugation, flow cytometric sorting and auto-and induced fluorescence; and 2) To determine the feasibility of invertebrate cell culture and microbial fermentation to produce bioactive metabolites. Specific Aim 2 would involve efforts to: a) culture metabolite-producing sponge-cells using methods developed in the principal investigator's laboratories modifying culture conditions to optimize cell growth and production of bioactive metabolites; and b) explore the use of fermentation techniques if bioactive compounds are found to be produced by associated microorganisms.